Objective: Adequate extravillous trophoblast (EVT) invasion plays a crucial role in the establishment of successful pregnancy. Insufficient trophoblast migration and invasion can result in defective placentation, which is associated with a number of clinical pathological conditions of pregnancy including spontaneous abortion and preeclampsia. Galectin-9 (Gal-9) has a wide variety of regulatory functions in innate and adaptive immunity during infection, tumor growth, and organ transplantation. Methods: We utilized immortalized human first-trimester EVT cells (HTR8/SVneo) for our functional study. We examined the effects of Gal-9 on viability, proliferation, and invasion of HTR8/SVneo cells, as well as on matrix metalloproteinase-2 (MMP-2) production in HTR8/SVneo cells. Furthermore, we observed the effects of different MAPK-signaling pathway inhibitors on the stimulatory functions of Gal-9 on HTR8/SVneo cells' invasion. Results: We verified the secretion of Gal-9 by trophoblasts and detected a correlation between low levels of Gal-9 and spontaneous abortion. Gal-9 promoted the invasion of HTR8/SVneo cells through its interaction with Tim-3, not CD44, and subsequently increased MMP-2 production. Blockade of p38 signaling pathway inhibited Gal-9 activities in HTR8/SVneo cells. Conclusion: Gal-9 promotes human trophoblast cell invasion through MMP-2 and p38 signaling pathway in a Tim-3-dependent manner.

Objective: To investigate whether kynurenine/aryl hydrocarbon receptor (AHR) affects the maternal-fetal tolerance by involving the differentiation of T helper 17 (Th17)/ regulatory T (Treg) cells, and to provide theoretical basic for new treatment of unexplained abortion. Methods: Flow cytometry (FCM) was used to detect the expression of AHR in peripheral/decidual CD4⁺ T, Treg, and Th17 cells. The effect of Kyn on the differentiation of peripheral/decidual naïve T-cells under Treg-/Th17-polarizing condition was detected by FCM; enzyme-linked immunosorbent assay was performed to examine the level of Kyn in villus and decidual tissues from normal pregnancy (NP) and unexplained abortion (UA). Student's t-test in the case of two groups or one-way ANOVA in multiple groups was used. Results: AHR expression in CD4⁺ T-cells was decreased in decidua versus blood in early pregnancy (P < 0.0001). Kyn could promote the differentiation of peripheral and decidual naïve T-cells to Th17 cells under Treg-polarizing conditions (P < 0.01). There was no statistical significance about the concentration of Kyn in decidual or villi tissues between NP and UA, and compared with NP, the expression of AHR in decidual CD4⁺ T-cells from UA was increased (P < 0.001). Conclusions: Kyn/AHR promotes Th17 and restricts Treg cells' differentiation, which is involved in maintaining the balance of Treg/Th17 cells at the maternal–fetal interface.

Objective: The aim is to study the effects of metformin on the expression of 70 kDa ribosomal protein S6 kinase (P70S6k), insulin receptor substrate 1 (IRS-1), and IRS-1Ser307 phosphorylation in human luteinized granulosa cells. Methods: Granulosa cells in the experimental group were cultured in M199 medium containing 0.1 mmol/L metformin for 24 h and those in control group were cultured in M199 medium. The expression levels of P70S6k and IRS-1 mRNA were detected by reverse-transcriptiom polymerase chain reaction (RT-PCR) and real-time PCR. P70S6k, IRS-1, p-ser307-IRS-1, and p-thr389-P70S6k protein expression levels were detected by immunofluorescence and western blotting. Results: P70S6k mRNA level was higher and IRS-1 was significantly lower in the experimental group than those in the control group. IRS-1 and p-ser307-IRS-1 were expressed in cell plasma, and P70S6k and p-thr389-P70S6k were expressed in cell nucleus. The results of Western blot analysis indicated that the expression levels of P70S6k, p-thr389-P70S6k, IRS-1, and p-ser307-IRS-1 proteins had significant difference between the experimental group and the control group. Compared to the control group, the relative intensity illustrated that the expression levels of P70S6K and p-thr389-P70S6k significantly increased in the experimental group; however, those of IRS-1 and p-ser307-IRS-1 proteins significantly decreased. Conclusion: Metformin can inhibit the P70S6k mRNA and protein expression levels in the granulosa cells and improve insulin sensitivity by regulating IRS-1 expression through Akt/P70S6k/IRS-1-dependent pathway.

Background: The aim of this study is to assess the effect of baicalein on chronic stress-mediated ovarian dysfunction in a mouse model. Methods: Forty female C57BL/6 mice were randomly divided into four groups as follows: the normal saline group (control, n = 10), the daily stress group (daily stress, n = 10), the baicalein group (baicalein, n = 10), and the daily stress + baicalein group (daily stress + baicalein, n = 10). For the daily stress model, we used a restricted stress model. Baicalein (10 mg/kg) was administered by gavage every day, and control mice received normal saline equivalently. Biopsy specimens were harvested after 4 weeks. Measurement of norepinephrine (NE) in serum was performed to assess the psychological stress level of the mice. In addition, histological changes of the uterus and ovaries and the levels of anti-Müllerian hormone (AMH) in serum were assessed to evaluate changes in ovarian function. To detect the underlying mechanisms of the amelioration of baicalein in chronic stress-mediated ovarian dysfunction, immunohistochemical methods, and quantitative real-time polymerase chain reaction were applied to determine the expression of gamma-aminobutyric acid (GABA) receptors. Results: Compared with values in the control group, serum NE concentrations were significantly increased (P < 0.001), AMH concentrations were markedly decreased (P < 0.01), the thickness of the endometrium was clearly reduced, and the percentage of atretic follicles was significantly increased in the daily stress group (P < 0.001), indicating that the chronic stress model was successfully established. In contrast, compared with values in the daily stress group, serum NE concentrations were significantly reduced (P < 0.001), AMH concentrations were significantly enhanced (P < 0.05), the thickness of the endometrium was clearly increased, and the percentage of atretic follicles was significantly reduced (P < 0.001) in the daily stress + baicalein group, indicating that baicalein clearly attenuated the ovarian dysfunction mediated by chronic stress. Moreover, the expression of the GABA_B2receptor in the daily stress group was significantly reduced (P < 0.01). In contrast, treatment with baicalein resulted in increased expression of the GABA_B2receptor (P < 0.01). Conclusions: Treatment with baicalein ameliorates the enhancing effect of chronic stress on ovarian dysfunction, and the mechanism can be attributed, in part, to the increased expression of the GABA_B2receptor.

Background: Reciprocal translocation (RCP) causes male infertility and female recurrent pregnancy loss. Male and female carriers have different responses to meiotic disturbances. Gender difference in outcomes of the RCP couples undergoing preimplantation genetic testing (PGT) is unknown. Methods: We conducted a retrospective analysis of 238 RCP couples (124 female and 114 male carriers) divided by gender of carrier from March 2014 to March 2017. Blastocysts were divided by day 5 and day 6. Females were divided into older (≥38 years) and younger (<38 years). Logistic regression was fitted for the relationship between gender of carriers and euploidy. Euploidy rate of each group, pregnancy rate, and live birth rate between different genders were analyzed. Results: The sperm live rate, forward motile sperm rate, and normal morphology rate of serum in male RCP group were significantly decreased. The euploidy rate was 30.30% in female group and 34.90% in male group (P = 0.131); 34.50% in day 5 group and 27.50% in day 6 group (P = 0.039); 33.40% in age <38 years group and 22.40% in age ≥38 years group (P = 0.063). Day 5 (odds ratio [OR] = 1.388, 95% confidence interval [CI] = 1.012–1.904; P = 0.042) and younger age (OR = 1.753, 95% CI = 0.97–3.17; P = 0.063) were associated with euploidy. The clinical pregnancy rate (37.90% vs. 41.20%), ongoing pregnancy rate (33.10% vs. 37.70%), and live birth rate (25.80% vs. 31.60%) per initiated were not significantly different in two gender groups. Conclusions: Although gender influence is not significant, couples with male carrier showed better clinical outcomes. The embryo growing rate and female age are important predictions estimating euploidy in RCP couples.

Background: To analyze the clinical outcomes of in vitro fertilization (IVF)/intracytoplasmic sperm injection treatments in women with a history of gestational trophoblastic disease (GTD). Methods: This retrospective study included 43 patients with a history of GTD as the study group and 43 matched patients as the control group. The patients in the study group were divided into two groups according to the therapy received. Patients in Subgroup A (n = 32) underwent uterine curettage treatment only. Patients in Subgroup B (n = 11) underwent uterine curettage combined with chemotherapy. The characteristics of ovarian stimulation and outcomes of embryos and pregnancy were compared. Results: In the first cycle, there was a higher number of retrieved oocytes and normal fertilized oocytes in the control group than those in the study group (9.2 vs. 6.2 and 6.0 vs. 4.0, respectively; P < 0.05); however, a similar mature oocyte rate (83.5% vs. 85.0%), normal fertilization rate (84.5% vs. 80.1%), number of good-quality embryos (1 vs. 2), and viable embryos (2 vs. 2) were found between the two groups (P > 0.05). There was no difference in the outcomes between Subgroup A and Subgroup B. There was a significant difference in thickness of the endometrium between the control group and study group (10.9 mm vs. 9.2 mm, respectively; P < 0.05). The biochemical pregnancy rate and ongoing pregnancy rate in the control group were significantly higher than those in the study group (51.4% vs. 31.7% and 37.8% vs. 18.3%, respectively; P < 0.05). In the study group, 28 (93.3%) patients had intrauterine adhesion (IUA) and 23 (76.7%) patients used an intrauterine device (IUD), which were both significantly higher than those in control group (P < 0.05). In addition, the rate of IUA in second-look hysteroscopy was lower than that in the first surgery in the study group (P < 0.05). Conclusions: Patients with a history of GTD can present with a similar normal fertilization rate and number of viable embryos. However, patients with a history of GTD may have a thinner endometrium and lower ongoing pregnancy rate. Hysteroscopy before frozen embryo transfer and usage of an IUD can improve the occurrence of IUA.

Background: Chronic inflammation has been reported as a triggering and driving factor in the pathogenesis of various diseases and is believed to be closely associated with tumor pathogenesis. In this study, we aimed to explore the changes of the inflammatory stress-related indexes during the menstrual period. Methods: Cytokines and tumor-associated carbohydrate antigens from 76 childbearing-age women during the follicular phase, luteal phase, and menstrual period were measured. Results: The expression of inflammatory indexes, such as platelets (PLT), lymphocytes (Lym), the percentage of Lym (Lym%), neutrophils (Neu), the percantage of Neu (Neu%), neutrophil-to-lymphocyte ratio (NLR), interleukin-6 (IL-6), and cancer antigen 125 (CA125), reached the highest level during the menstrual period. Conclusions: Female genital organs are under inflammatory stress during menstruation, which hints that the changes of the inflammatory state of the body play an important role in the pathogenesis of diseases.

Zearalenone (ZEA) is produced by Fusarium species and a common contaminant in food. ZEA and its metabolites, α- and β-zearalenol, α- and β-zearalanol, and zearalanone, are mycoestrogens that can interfere with estrogen signaling. High levels of mycoestrogens reduced female fertility in farm animals and rodents, in which adverse effects of mycoestrogens on major events in female reproduction, including ovarian folliculogenesis, ovulation, ovarian steroidogenesis, fertilization, preimplantation embryo development and transport, embryo implantation, placentation, parturition, and lactation, have been reported in different experimental settings. Here, we review the in vivo effects of mycoestrogens on the main events in female reproduction.

The recent progress in the association of cumulus cells with immune functions is a largely ignored area. With over 350 million new sexually transmitted infections occurring annually in adults of reproductive age, we feel the need to explore more about how the cumulus cells defend themselves and protect the oocytes during the development through the ovulation period. Application of assisted reproductive technologies allows scientists to study and better understand cumulus cells. There are still many immune factors to be taken into consideration to optimize the oocyte quality besides ovarian stimulations. The objective of this review is to summarize the key elements of cumulus cells and their association with the immune function.